{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T04:36:50Z","timestamp":1790743010616,"version":"4.1.0"},"reference-count":50,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2020,11,26]],"date-time":"2020-11-26T00:00:00Z","timestamp":1606348800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 EY023598"],"award-info":[{"award-number":["R01 EY023598"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 EY011915"],"award-info":[{"award-number":["R01 EY011915"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["Core Grant for Vision Research EY002520"],"award-info":[{"award-number":["Core Grant for Vision Research EY002520"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Lions Foundation for Sight","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"Research to Prevent Blindness Unrestricted","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"Oshman Foundation","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"William Stamps Farish Fund","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["Young Scientists Grant 81400380"],"award-info":[{"award-number":["Young Scientists Grant 81400380"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["Young Scientists Grant 82000862"],"award-info":[{"award-number":["Young Scientists Grant 82000862"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012226","name":"Fundamental Research Funds for the Central Universities","doi-asserted-by":"publisher","award":["XJJ2014076"],"award-info":[{"award-number":["XJJ2014076"]}],"id":[{"id":"10.13039\/501100012226","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Inflammation is the main pathophysiology of dry eye, characterized by tear film instability and hyperosmolarity. The aim of this study was to investigate the association of inflammation and cellular autophagy using an in vitro dry eye model with primary cultured human corneal epithelial cells (HCECs). Primary HCECs cultured with fresh limbal explants from donors were switched to a hyperosmotic medium (450 mOsM) by adding sodium chloride into the culture medium. We observed the stimulated inflammatory mediators, TNF-\u03b1, IL-1\u03b2, IL-6 and IL-8, as well as the increased expression of autophagy related genes, Ulk1, Beclin1, Atg5 and LC3B, as evaluated by RT-qPCR and ELISA. The immunofluorescent staining of LC3B and Western blotting revealed the activated autophagosome formation and autophagic flux, as evidenced by the increased LC3B autophagic cells with activated Beclin1, Atg5, Atg7 and LC3B proteins, and the decreased levels of P62 protein in HCECs. Interestingly, the autophagy activation was later at 24 h than inflammation induced at 4 h in HCECs exposed to 450 mOsM. Furthermore, application of rapamycin enhanced autophagy activation also reduced the inflammatory mediators and restored cell viability in HCECs exposed to the hyperosmotic medium. Our findings for the first time demonstrate that the autophagy activation is a late phase response to hyperosmotic stress, and is enhanced by rapamycin, which protects HCECs by suppressing inflammation and promoting cells survival, suggesting a new therapeutic potential to treat dry eye diseases.<\/jats:p>","DOI":"10.3390\/ijms21238966","type":"journal-article","created":{"date-parts":[[2020,11,26]],"date-time":"2020-11-26T09:04:15Z","timestamp":1606381455000},"page":"8966","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":55,"title":["Autophagy Activation Protects Ocular Surface from Inflammation in a Dry Eye Model In Vitro"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8818-0549","authenticated-orcid":false,"given":"Zhao","family":"Liu","sequence":"first","affiliation":[{"name":"Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USA"},{"name":"Department of Ophthalmology, the First Affiliated Hospital of Xi\u2019an Jiaotong University, Xi\u2019an 710061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6890-7384","authenticated-orcid":false,"given":"Ding","family":"Chen","sequence":"additional","affiliation":[{"name":"Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USA"},{"name":"School of Ophthalmology &amp; Optometry, Wenzhou Medical University, Wenzhou 325027, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Chen","sequence":"additional","affiliation":[{"name":"Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USA"},{"name":"School of Ophthalmology &amp; Optometry, Wenzhou Medical University, Wenzhou 325027, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fang","family":"Bian","sequence":"additional","affiliation":[{"name":"Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ning","family":"Gao","sequence":"additional","affiliation":[{"name":"Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USA"},{"name":"Department of Ophthalmology, the First Affiliated Hospital of Xi\u2019an Jiaotong University, Xi\u2019an 710061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinmiao","family":"Li","sequence":"additional","affiliation":[{"name":"Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen C.","family":"Pflugfelder","sequence":"additional","affiliation":[{"name":"Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"De-Quan","family":"Li","sequence":"additional","affiliation":[{"name":"Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,11,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1016\/j.jtos.2017.05.003","article-title":"TFOS DEWS II Epidemiology Report","volume":"15","author":"Stapleton","year":"2017","journal-title":"Ocul. Surf."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1016\/j.jtos.2017.05.008","article-title":"TFOS DEWS II Definition and Classification Report","volume":"15","author":"Craig","year":"2017","journal-title":"Ocul. Surf"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1117","DOI":"10.1001\/jamaophthalmol.2015.2326","article-title":"Effects of azithromycin on gene expression profiles of proinflammatory and anti-inflammatory mediators in the eyelid margin and conjunctiva of patients with meibomian gland disease","volume":"133","author":"Zhang","year":"2015","journal-title":"JAMA Ophthalmol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.jaut.2017.02.006","article-title":"Mitochondrial DNA oxidation induces imbalanced activity of NLRP3\/NLRP6 inflammasomes by activation of caspase-8 and BRCC36 in dry eye","volume":"80","author":"Chi","year":"2017","journal-title":"J. Autoimmun."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1016\/j.jtos.2017.05.001","article-title":"TFOS DEWS II Diagnostic Methodology report","volume":"15","author":"Wolffsohn","year":"2017","journal-title":"Ocul Surf"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"7545","DOI":"10.1167\/iovs.15-17627","article-title":"Aqueous Tear Deficiency Increases Conjunctival Interferon-gamma (IFN-gamma) Expression and Goblet Cell Loss","volume":"56","author":"Pflugfelder","year":"2015","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"900","DOI":"10.1016\/j.ajo.2011.08.023","article-title":"Tear dysfunction and the cornea: LXVIII Edward Jackson Memorial Lecture","volume":"152","author":"Pflugfelder","year":"2011","journal-title":"Am. J. Ophthalmol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1016\/j.jtos.2013.07.003","article-title":"Role of hyperosmolarity in the pathogenesis and management of dry eye disease: Proceedings of the OCEAN group meeting","volume":"11","author":"Baudouin","year":"2013","journal-title":"Ocul. Surf."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"S4","DOI":"10.1016\/j.ophtha.2017.07.010","article-title":"The Pathophysiology of Dry Eye Disease: What We Know and Future Directions for Research","volume":"124","author":"Pflugfelder","year":"2017","journal-title":"Ophthalmology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1097\/01.ICL.0000162759.79740.46","article-title":"Hyperosmolar saline is a proinflammatory stress on the mouse ocular surface","volume":"31","author":"Luo","year":"2005","journal-title":"Eye Contact Lens"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.exer.2013.11.011","article-title":"Topical interferon-gamma neutralization prevents conjunctival goblet cell loss in experimental murine dry eye","volume":"118","author":"Zhang","year":"2014","journal-title":"Exp Eye Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6287","DOI":"10.1167\/iovs.13-12081","article-title":"Efficacy of osmoprotectants on prevention and treatment of murine dry eye","volume":"54","author":"Chen","year":"2013","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4302","DOI":"10.1167\/iovs.04-0299","article-title":"Stimulation of matrix metalloproteinases by hyperosmolarity via a JNK pathway in human corneal epithelial cells","volume":"45","author":"Li","year":"2004","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1016\/j.exer.2005.08.019","article-title":"JNK and ERK MAP kinases mediate induction of IL-1beta, TNF-alpha and IL-8 following hyperosmolar stress in human limbal epithelial cells","volume":"82","author":"Li","year":"2006","journal-title":"Exp. Eye Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"19408","DOI":"10.1038\/srep19408","article-title":"Blueberry Component Pterostilbene Protects Corneal Epithelial Cells from Inflammation via Anti-oxidative Pathway","volume":"6","author":"Li","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"657","DOI":"10.3109\/02713683.2014.957776","article-title":"Effects of L-carnitine, erythritol and betaine on pro-inflammatory markers in primary human corneal epithelial cells exposed to hyperosmotic stress","volume":"40","author":"Hua","year":"2015","journal-title":"Curr. Eye Res."},{"key":"ref_17","first-page":"1243","article-title":"Osmoprotectants suppress the production and activity of matrix metalloproteinases induced by hyperosmolarity in primary human corneal epithelial cells","volume":"20","author":"Deng","year":"2014","journal-title":"Mol. Vis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1016\/j.jtos.2017.05.006","article-title":"TFOS DEWS II Management and Therapy Report","volume":"15","author":"Jones","year":"2017","journal-title":"Ocul. Surf."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1016\/j.preteyeres.2016.08.001","article-title":"Autophagy in the eye: Development, degeneration, and aging","volume":"55","author":"Boya","year":"2016","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/j.ceb.2010.10.009","article-title":"Chaperone-mediated autophagy in protein quality control","volume":"23","author":"Arias","year":"2011","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1038\/nrm4024","article-title":"Autophagy at the crossroads of catabolism and anabolism","volume":"16","author":"Kaur","year":"2015","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1038\/nature07383","article-title":"Loss of the autophagy protein Atg16L1 enhances endotoxin-induced IL-1beta production","volume":"456","author":"Saitoh","year":"2008","journal-title":"Nature"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1016\/j.cell.2016.05.051","article-title":"Autophagy, Inflammation, and Immunity: A Troika Governing Cancer and Its Treatment","volume":"166","author":"Zhong","year":"2016","journal-title":"Cell"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1186\/s40169-017-0154-5","article-title":"Autophagy and inflammation","volume":"6","author":"Qian","year":"2017","journal-title":"Clin. Transl. Med."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Crisan, T.O., Plantinga, T.S., van de Veerdonk, F.L., Farcas, M.F., Stoffels, M., Kullberg, B.J., van der Meer, J.W., Joosten, L.A., and Netea, M.G. (2011). Inflammasome-independent modulation of cytokine response by autophagy in human cells. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0018666"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"884","DOI":"10.7150\/ijbs.15194","article-title":"Interactions between Autophagy and Inhibitory Cytokines","volume":"12","author":"Wu","year":"2016","journal-title":"Int. J. Biol. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"C73","DOI":"10.1152\/ajpcell.00110.2016","article-title":"Cigarette smoke-induced autophagy impairment accelerates lung aging, COPD-emphysema exacerbations and pathogenesis","volume":"314","author":"Vij","year":"2016","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/s10565-016-9374-5","article-title":"Autophagy-from molecular mechanisms to clinical relevance","volume":"33","author":"Lippai","year":"2017","journal-title":"Cell Biol. Toxicol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1146","DOI":"10.4161\/auto.20520","article-title":"LC3 as a potential therapeutic target in hypoxia-induced pulmonary hypertension","volume":"8","author":"Lahm","year":"2012","journal-title":"Autophagy"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1002\/mgg3.258","article-title":"CFTR founder mutation causes protein trafficking defects in Chinese patients with cystic fibrosis","volume":"5","author":"Leung","year":"2017","journal-title":"Mol. Genet. Genom. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1097\/MIB.0000000000000994","article-title":"The Crohn\u2019s and Colitis Foundation of America Survey of Inflammatory Bowel Disease Patient Health Care Access","volume":"23","author":"Rubin","year":"2017","journal-title":"Inflamm. Bowel Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"17280","DOI":"10.1038\/s41598-017-17128-0","article-title":"Elevation of autophagy markers in Sjogren syndrome dry eye","volume":"7","author":"Byun","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1016\/j.exer.2019.04.023","article-title":"Corneal autophagy and ocular surface inflammation: A new perspective in dry eye","volume":"184","author":"Ma","year":"2019","journal-title":"Exp. Eye Res."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Deng, R., Hua, X., Li, J., Chi, W., Zhang, Z., Lu, F., Zhang, L., Pflugfelder, S.C., and Li, D.Q. (2015). Oxidative stress markers induced by hyperosmolarity in primary human corneal epithelial cells. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0126561"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1097\/ICO.0b013e318030d259","article-title":"Hyperosmolarity-induced apoptosis in human corneal epithelial cells is mediated by cytochrome c and MAPK pathways","volume":"26","author":"Luo","year":"2007","journal-title":"Cornea"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1167\/iovs.07-0569","article-title":"Hyperosmolarity-induced cornification of human corneal epithelial cells is regulated by JNK MAPK","volume":"49","author":"Chen","year":"2008","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1016\/j.jtos.2019.01.008","article-title":"Autophagy in corneal health and disease: A concise review","volume":"17","author":"Martin","year":"2019","journal-title":"Ocul. Surf."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1007\/978-1-4939-8873-0_26","article-title":"In Vitro Screening Platforms for Identifying Autophagy Modulators in Mammalian Cells","volume":"1880","author":"Seranova","year":"2019","journal-title":"Methods Mol. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1038\/aps.2012.184","article-title":"Why should autophagic flux be assessed?","volume":"34","author":"Zhang","year":"2013","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1111\/imr.12286","article-title":"Initiation and perpetuation of NLRP3 inflammasome activation and assembly","volume":"265","author":"Elliott","year":"2015","journal-title":"Immunol. Rev."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"896","DOI":"10.1016\/j.cell.2015.12.057","article-title":"NF-kappaB Restricts Inflammasome Activation via Elimination of Damaged Mitochondria","volume":"164","author":"Zhong","year":"2016","journal-title":"Cell"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.lfs.2017.12.001","article-title":"Diabetic retinopathy pathogenesis and the ameliorating effects of melatonin; involvement of autophagy, inflammation and oxidative stress","volume":"193","author":"Dehdashtian","year":"2018","journal-title":"Life Sci."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1038\/nature17039","article-title":"Reparative inflammation takes charge of tissue regeneration","volume":"529","author":"Karin","year":"2016","journal-title":"Nature"},{"key":"ref_44","first-page":"37","article-title":"Matrix metalloproteinase-9 in the pathophysiology and diagnosis of dry eye syndrome","volume":"4","author":"Pflugfelder","year":"2017","journal-title":"Met. Med."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"5503","DOI":"10.1167\/iovs.14-16247","article-title":"Protective Effects of L-Carnitine Against Oxidative Injury by Hyperosmolarity in Human Corneal Epithelial Cells","volume":"56","author":"Hua","year":"2015","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Li, Y., Liu, H., Zeng, W., and Wei, J. (2017). Edaravone protects against hyperosmolarity-induced oxidative stress and apoptosis in primary human corneal epithelial cells. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0174437"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.exer.2004.02.015","article-title":"Phenotypic characterization of human corneal epithelial cells expanded ex vivo from limbal explant and single cell cultures","volume":"79","author":"Kim","year":"2004","journal-title":"Exp. Eye Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"5623","DOI":"10.1167\/iovs.09-4992","article-title":"Suppressive effects of azithromycin on zymosan-induced production of proinflammatory mediators by human corneal epithelial cells","volume":"51","author":"Li","year":"2010","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1634\/stemcells.22-3-355","article-title":"Characterization of putative stem cell phenotype in human limbal epithelia","volume":"22","author":"Chen","year":"2004","journal-title":"Stem Cells"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"7336","DOI":"10.1016\/j.biomaterials.2012.06.080","article-title":"Using genipin-crosslinked acellular porcine corneal stroma for cosmetic corneal lens implants","volume":"33","author":"Liu","year":"2012","journal-title":"Biomaterials"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/23\/8966\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:37:28Z","timestamp":1760179048000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/23\/8966"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,26]]},"references-count":50,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2020,12]]}},"alternative-id":["ijms21238966"],"URL":"https:\/\/doi.org\/10.3390\/ijms21238966","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,11,26]]}}}